Diagnosis and treatment of infection involving killer t follicular helper cells, methods of preparation, and uses thereof

Abstract

Recurrent tonsillitis disease (RT) is a common indication for pediatric tonsillectomy, the most frequent childhood surgery. It is unknown why some children develop RT. The present disclosure demonstrates that RT tonsils exhibit significantly smaller germinal centers than non-RT tonsils, concomitant with a bias against Group A Streptococcus (GAS)-specific germinal center follicular helper CD4+ T cells (GC Tfh), and significantly reduced antibodies to the GAS virulence factor SpeA. The present disclosure also shows a significant immunogenetic component to this disease, with the identification of ‘at risk’ and ‘protective’ HLA alleles for RT. Finally, the present disclosure identifies a new cell type, granzyme B+GC Tfh cells, which are activated by SpeA, are significantly more abundant in RT GC Tfh cells, and have the capacity to kill B cells, thus, providing a window into the immunology and genetics of a classic childhood disease and identifies a new type of pathogenic T cell.

The invention claimed is: 
     
       1. A method for treatment of a subject for tonsillitis, recurrent tonsillitis, or strep throat, the method comprising:
 processing a biological sample from a subject, wherein the biological sample is a tonsillar tissue or a lymph node tissue sample, 
 the sample being suspected of including Granzyme B+germinal center T follicular helper cells (Granzyme B+GC Tfh cells), by measuring an amount of the Granzyme B+GC Tfh cells which are specific for or responsive to Streptococcal pyrogenic exotoxin A (SpeA), and 
 comparing the measured amount to a reference amount obtained from a subject without strep throat or tonsillitis, wherein if the measured amount of Granzyme B+GC Tfh cells is greater than the amount in the reference amount it is indicative that the subject has, is at risk of having, or is need of treatment for tonsillitis, recurrent tonsillitis, or strep throat and 
 treating the subject if the measured amount of Granzyme B+GC Tfh cells is more than the amount in the reference amount based on the determining step with an agent that decreases the Granzyme B+GC Tfh cells in the subject. 
 
     
     
       2. The method of  claim 1 , wherein measuring the Granzyme B+GC Tfh cells specific for or responsive to SpeA is performed using an antigen-specific CD4 T cell activation induced marker (AIM) assay. 
     
     
       3. The method of  claim 2 , wherein the antigen-specific CD4 T cell AIM assay comprises detecting CD25, Ox40 and PD-L1. 
     
     
       4. The method of  claim 1 , wherein treating the subject comprises administering a vaccination comprising SpeA, or an SpeA peptide for tonsillitis, recurrent tonsillitis, or strep throat. 
     
     
       5. The method of  claim 1 , wherein the reference amount is obtained by processing a control sample which is a sample from the subject prior to the treatment, and wherein an increase in the measured amount of Granzyme B+GC Tfh cells compared to the reference amount is indicative that the treatment is effective, or wherein the measuring Granzyme B+GC Tfh cells specific for or responsive to SpeA is performed using an activation induced marker (AIM) assay. 
     
     
       6. The method of  claim 1 , wherein the method comprises providing an agent for treatment of the subject for tonsillitis, recurrent tonsillitis, or strep throat, wherein the agent is selected from an immunization with SpeA, or an SpeA peptide, or a peptide, protein, recombinant protein, recombinant peptides, antibody, small molecule, ligand mimetic, or a nucleic acid that modulates, reduces, inhibits, decreases or blocks SpeA. 
     
     
       7. A method for evaluating a condition status in a subject, the condition being a disease or disorder associated with impaired germinal centers, the method comprising:
 a. providing a biological sample, wherein the biological sample is a tonsillar tissue or a lymph node tissue sample, from said subject, the sample being suspected of including Granzyme B+germinal center T follicular helper cells (Granzyme B+GC Tfh cells); 
 b. processing the sample to determine a concentration, activation, differentiation, proliferation or activity level of said Granzyme B+GC Tfh cells in said sample; 
 c. comparing the concentration, activation, differentiation, proliferation or activity level to a reference level obtained from a subject without strep throat, tonsillitis or an autoimmune disease or an average measurement or value gathered from a population of healthy individuals that do not have strep throat, tonsillitis or an autoimmune disease; 
 d. evaluating the condition status based on at least the comparison in step (c), the condition being associated with impaired germinal centers, wherein the condition status is the disease or disorder associated with impaired germinal centers is tonsillitis, strep throat, recurrent tonsillitis, or the autoimmune disease in the subject based on Granzyme B+GC Tfh cells in said sample compared to the reference level; and 
 e. further comprising treating said subject at least based on said comparison by administering an agent that modulates, increases, enhances, elicits, stimulates, promotes activation, differentiation, proliferation, number or activity of Granzyme B+killer GC Tfh cells, wherein the agent stimulates activation, differentiation, proliferation, number or activity of Granzyme B+killer GC Tfh cells so as to modulate the concentration of said Granzyme B+killer GC Tfh cells in said subject to treat the autoimmune disease. 
 
     
     
       8. The method of  claim 7 , further comprising the step of determining response or resistance to treatment for a disease or disorder associated with impaired germinal centers in a subject undergoing treatment for a disease or disorder associated with impaired germinal centers. 
     
     
       9. The method of  claim 7 , further comprising treating the autoimmune disease, the method comprising administering an agent to said subject for modulating, increasing, enhancing, eliciting, stimulating or promoting activation, differentiation, proliferation, number or activity of Granzyme B+GC Tfh cells. 
     
     
       10. The method of  claim 9 , further comprising administering to the subject an effective amount of a purified Granzyme B- germinal center T follicular helper cell population sufficient to treat the tonsillitis, strep throat, or recurrent tonsillitis. 
     
     
       11. The method of  claim 7 , wherein the autoimmune disease is an autoantibody associated autoimmune disease. 
     
     
       12. The method of  claim 10 , further comprising treating unstimulated GC Tfh cells to have modified gene expression, wherein the modified gene expression comprises expression of Granzyme B and at least one of: increased expression of at least one of: PRDM1 (BLIMP1), decreased expression of BCL1, increased expression of ICOS, increased expression of GZMB, decreased expression of CD28, increased expression of CTLA4, increased expression of EOMES, or increased expression of TBX21 (T-bet), to make Granzyme B+GC Tfh cells. 
     
     
       13. The method of  claim 1 , wherein treating the subject for tonsillitis, recurrent tonsillitis, or strep throat comprises administering an agent to the subject that: a) increases SpeA specific Granzyme B- germinal center T follicular helper cells, b) increases antibodies against SpeA, c) modulates, reduces, inhibits, decreases or blocks SpeA in an amount sufficient to treat tonsillitis or strep throat in the subject, or d) modulates, reduces, inhibits, decreases or blocks differentiation or activity of the Granzyme B+killer GC Tfh cells. 
     
     
       14. The method of  claim 4 , wherein the vaccination for tonsillitis or strep throat further comprises an adjuvant.